尿激酶型纤溶酶原激活因子体外诱导小鼠精子趋化运动的分子机制探讨.docVIP

  尿激酶型纤溶酶原激活因子体外诱导小鼠 精子趋化运动的分子机制探讨# 丁晓芳1，李红钢2，官黄涛2* 5 10 15 20 25 30 35 40 45 （1. 华中科技大学，附属协和医院生殖医学中心，武汉 430022； 2. 华中科技大学，同济医学院计划生育研究所，武汉 430030） 摘要：目的 探讨尿激酶型纤溶酶原激活因子体外诱导小鼠精子趋化运动的作用机制。方法 uPA 体外诱导获能小鼠精子产生趋化运动，收集 uPA 作用不同时间（0、5、30、60 min） 后产生趋化运动的精子，设为实验组；以相同时间段的获能精子作为对照组。利用激光共聚 焦显微镜检测精子胞内 Ca2+-Fluo3-AM 荧光强度；同时利用 Western Blot 检测各组各时间点 精子胞浆内磷酸化 ERK1/2 蛋白水平。结果 在第 5、30 min 时捕捉的精子胞内 Fluo3-AM 荧 光最强，较对照组同时间点相比显著增强（P 0.05）；与相同时间点的对照组比较， pERK1/ERK1 比值在 5 min、30 min 和 60 min 时升高显著（P0.05），pERK2/ERK2 比值在 30 min 和 60min 时增加有显著差异（P0.05）。结论 uPA 体外诱导小鼠精子产生趋化运动 的分子机制，与增加精子[Ca2+]i 和磷酸化 ERK1/2 蛋白水平有关。 关键词：尿激酶型纤溶酶原激活因子；精子；趋化性；钙离子；ERK 称 中图分类号：R339.2+1 Study of molecular mechanism of mouse spermatozoa chemotactic movement induced by Urokinase-Type Plasminogen Activator in vitro DING Xiaofang1, LI Honggang2, GUAN Huangtao2 (1. Center of Reproductive Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, WuHan 430022; 2. Family Planning Rsearch Institute, Tobngji Medical College, Huazhong University of Science and Technology, WuHan 430030) Abstract: Objective: To study the molecular mechanism of mouse spermatozoa chemotactic movement induced by Urokinase-Type plasminogene activator (uPA) in vitro. Methods: chemotaxis of mouse capacitated-sperm was induced by uPA in vitro. The chemotactic sperm were divided into 4 test groups according to its different incubating time with uPA (0，5，30， 60min respectively). The capacitated-sperm without uPA guide were designed as control groups and collected at the same time points. The fluorescence intensity of intracellular [Ca2+]i and the levels of phosphorylated ERK1/2 proteins in sperm in each group were determined by confocal microscopy and Western Blot respectively. Results: the mean fluorescence intensities of sperm [Ca2+]i in test groups were significantly increased than that of in the control groups at 5 and 30 min (P0.05), and the levels of phosphorylated ERK1 and ERK2 proteins were also increased ob